Expression of pH-sensitive green fluorescent protein in Arabidopsis thaliana

Plant Cell Environ. 2001 May;24(5):557-63. doi: 10.1046/j.1365-3040.2001.00703.x.

Abstract

This is the first report on using green fluorescent protein (GFP) as a pH reporter in plants. Proton fluxes and pH regulation play important roles in plant cellular activity and therefore, it would be extremely helpful to have a plant gene reporter system for rapid, non-invasive visualization of intracellular pH changes. In order to develop such a system, we constructed three vectors for transient and stable transformation of plant cells with a pH-sensitive derivative of green fluorescent protein. Using these vectors, transgenic Arabidopsis thaliana and tobacco plants were produced. Here the application of pH-sensitive GFP technology in plants is described and, for the first time, the visualization of pH gradients between different developmental compartments in intact whole-root tissues of A. thaliana is reported. The utility of pH-sensitive GFP in revealing rapid, environmentally induced changes in cytoplasmic pH in roots is also demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabidopsis / chemistry
  • Arabidopsis / cytology
  • Arabidopsis / genetics*
  • Cytoplasm / chemistry*
  • Gene Expression Regulation, Plant
  • Genes, Plant
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins
  • Hydrogen-Ion Concentration
  • Luminescent Proteins / genetics*
  • Microscopy, Fluorescence
  • Plant Roots / chemistry
  • Plant Roots / cytology*
  • Plant Roots / genetics
  • Plants, Genetically Modified
  • Plasmids / genetics
  • Protons*
  • Tobacco / genetics

Substances

  • Luminescent Proteins
  • Protons
  • Green Fluorescent Proteins